Proteomics/Genomics of Opiate Analgesia and Addiction

简介（由申请人提供）：吗啡是一种高效的阿片类镇痛药，是现代医学中一种有价值的止痛药。然而，长期使用吗啡的缺点是产生成瘾、耐受性和依赖性。理想情况下，设计有效镇痛的治疗方法将是有利的，它缺乏吗啡和相关阿片类药物的成瘾性。考虑到这一目标，有必要了解与吗啡耐受性和依赖性相比，吗啡诱导镇痛的生化途径。当新的创新技术应用于阐明具体的研究问题时，往往会出现科学进步。因此，利用蛋白质组学和基因组学的最新技术进行先导性创新研究的R21申请将阐明与吗啡诱导的镇痛耐受或依赖相关的分子成分。值得注意的是，β -抑制素敲除小鼠在吗啡诱导的行为中表现出明显的变化，相应的镇痛增强，缺乏耐受性，依赖性没有变化。这些发现导致吗啡作用的这三个参数有不同的生化途径的假设。因此，本可行性项目的目标将是利用蛋白质组学和基因组学来比较慢性吗啡治疗期间β -抑制素敲除小鼠和野生型小鼠的调节分子成分，作为阐明吗啡诱导镇痛与耐受或依赖相比的不同生化途径的手段。具体目的是：(1)与野生型小鼠相比，β -抑制素敲除小鼠在慢性吗啡期间大脑中调节的蛋白质和肽的蛋白质组学评估；(2)使用独特的神经特异性小鼠cDNA微阵列对β -抑制素敲除小鼠和野生型小鼠在慢性吗啡期间的差异基因表达进行基因组评估。这些研究将利用最先进的蛋白质组学和质谱分析设备，以及神经特异性小鼠cDNA微阵列基因组学。对调控基因产物进行基因家族一级序列同源性分析，预测其生物学功能。在β -抑制素敲除小鼠和野生型小鼠中比较受调控的基因产物将允许分配受调控的蛋白质和基因，以确定参与吗啡镇痛、耐受或依赖的候选生化途径。阐明不同的途径将是重要的，因为它可以允许未来设计阿片类药物方案，提供有效的镇痛，没有成瘾的耐受性和依赖性。

DESCRIPTION (provided by applicant): Morphine is a highly effective opiate analgesic, and is a valuable painkiller in modern medicine. However, the disadvantage of chronic morphine administration is the development of addiction with tolerance and dependence. Ideally, it would be advantageous to design therapeutics for effective analgesia, which lacks the addictive liabilities of morphine and related opiates. With this goal in mind, it is necessary to understand the biochemical pathways that mediate morphine-induced analgesia compared to tolerance, and dependence. Scientific advances often occur when new innovative technologies are applied towards elucidating specific research issues. Therefore, this R21 application for pilot, innovative studies utilizing state-of-the-art technologies in both proteomics and genomics, will elucidate molecular components involved in morphine-induced analgesia compared to tolerance or dependence. Notably, beta-arrestin knockout mice show distinct changes in morphine-induced actions corresponding to enhanced analgesia, absence of tolerance, and no change in dependence. These findings lead to the hypothesis of distinct biochemical pathways for these three parameters of morphine actions. Therefore, the goal of this feasibility project will be to utilize both proteomics and genomics to compare regulated molecular components in beta-arrestin knockout mice and wild-type mice during chronic morphine treatment, as a means to elucidate distinct biochemical pathways for morphine-induced analgesia compared to tolerance or dependence. The specific aims are (1) proteomic evaluation of regulated proteins and peptides during chronic morphine in brains of beta-arrestin knockout mice compared to wild-type mice, and (2) genomic evaluation of differential gene expression during chronic morphine using unique neural-specific mouse cDNA microarrays in beta-arrestin knockout and wild-type mice. These studies will utilize state-of-the-art facilities for proteomics and mass spectrometry, and genomics with neural-specific mouse cDNA microarrays. Regulated gene products will be analyzed for primary sequence homology to gene families to predict their biological functions. Comparison of regulated gene products in beta-arrestin knockout and wild-type mice will allow assignment of regulated proteins and genes to candidate biochemical pathways involved in morphine analgesia, tolerance, or dependence. Elucidation of distinct pathways will be significant, since it could allow future design of opiate drug regimens that provide effective analgesia, without the tolerance and dependence of addiction.